DRP1 Antibody | Affinity Biosciences

製品:	DRP1 Antibody
カタログ:	DF7037
タンパク質の説明:	Rabbit polyclonal antibody to DRP1
アプリケーション:	WB IHC IF/ICC
反応性:	Human, Mouse, Rat
予測:	Pig, Zebrafish, Bovine, Horse, Sheep, Rabbit, Dog, Chicken
分子量:	79kDa; 82kD(Calculated).
ユニプロット:	O00429
RRID:	AB_2838993

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サイズ	価格	在庫状況
100ul	$280	在庫あり
200ul	$350	在庫あり

プロトコル

製品説明

ソース:

Rabbit

アプリケーション:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

反応性:

Human,Mouse,Rat

予測:

Pig(100%), Zebrafish(83%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(83%)

クローナリティ:

Polyclonal

特異性:

DRP1 Antibody detects endogenous levels of total DRP1.

RRID:

AB_2838993
引用形式: Affinity Biosciences Cat# DF7037, RRID:AB_2838993.

コンジュゲート:

Unconjugated.

精製:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

保存:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

別名:

折りたたみ／展開

DLP1; dnm1l; DNM1L_HUMAN; Dnm1p/Vps1p-like protein; dnml1; DRP1; DVLP; Dymple; Dynamin 1 like; Dynamin family member proline-rich carboxyl-terminal domain less; Dynamin like protein; Dynamin related protein 1; Dynamin-1-like protein; Dynamin-like protein 4; Dynamin-like protein; Dynamin-like protein IV; Dynamin-related protein 1; DYNIV 11; EMPF; EMPF1; FLJ41912; HdynIV; VPS1;

免疫原

免疫原:

Uniprot:

O00429(DNM1L_HUMAN) >>Visit HPA database.

遺伝子(ID):

DNM1L(10059)

発現特異性:

O00429 DNM1L_HUMAN:

Ubiquitously expressed with highest levels found in skeletal muscles, heart, kidney and brain. Isoform 1 is brain-specific. Isoform 2 and isoform 3 are predominantly expressed in testis and skeletal muscles respectively. Isoform 4 is weakly expressed in brain, heart and kidney. Isoform 5 is dominantly expressed in liver, heart and kidney. Isoform 6 is expressed in neurons.

タンパク質の説明:

Dynamin-related protein 1 (DRP1) is a member of the dynamin superfamily of GTPases. Members of this family have diverse cellular functions including vesicle scission, organelle fission, viral resistance, and intracellular trafficking (reviewed in 1). DRP1 affects mitochondrial morphology and is important in mitochondrial and peroxisomal fission in mammalian cells (2-5). The yeast ortholog of DRP1 clusters into a spiral-shaped structure on the mitochondrial membrane at the site of fission (reviewed in 6), and this structure is likely conserved in mammalian cells (3). The division of the mitochondria, which is required for apoptosis, as well as normal cell growth and development is controlled, in part, by the phosphorylation of DRP1 at Ser616 by Cdk1/cyclin B and at Ser637 by protein kinase A (PKA) (reviewed in 6). When phosphorylated at Ser616, DRP1 stimulates mitochondrial fission during mitosis. Conversely, fission is inhibited when DRP1 is phosphorylated at Ser637 (reviewed in 6). Dephosphorylation at Ser637 by calcineurin reverses this inhibition (7). In addition to phosphorylation, sumoylation of DRP1 is also an enhancer of mitochondrial fission (8). Balancing fission and fusion events is essential for proper mitochondrial function. Research studies have demonstrated mitochondrial defects in a variety of neurodegenerative diseases including Alzheimer’s disease, Parkinson’s disease, and Huntington’s disease (reviewed in 6).

タンパク質配列:

O00429 DNM1L_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MEALIPVINKLQDVFNTVGADIIQLPQIVVVGTQSSGKSSVLESLVGRDLLPRGTGIVTRRPLILQLVHVSQEDKRKTTGEENGVEAEEWGKFLHTKNKLYTDFDEIRQEIENETERISGNNKGVSPEPIHLKIFSPNVVNLTLVDLPGMTKVPVGDQPKDIELQIRELILRFISNPNSIILAVTAANTDMATSEALKISREVDPDGRRTLAVITKLDLMDAGTDAMDVLMGRVIPVKLGIIGVVNRSQLDINNKKSVTDSIRDEYAFLQKKYPSLANRNGTKYLARTLNRLLMHHIRDCLPELKTRINVLAAQYQSLLNSYGEPVDDKSATLLQLITKFATEYCNTIEGTAKYIETSELCGGARICYIFHETFGRTLESVDPLGGLNTIDILTAIRNATGPRPALFVPEVSFELLVKRQIKRLEEPSLRCVELVHEEMQRIIQHCSNYSTQELLRFPKLHDAIVEVVTCLLRKRLPVTNEMVHNLVAIELAYINTKHPDFADACGLMNNNIEEQRRNRLARELPSAVSRDKSSKVPSALAPASQEPSPAASAEADGKLIQDSRRETKNVASGGGGVGDGVQEPTTGNWRGMLKTSKAEELLAEEKSKPIPIMPASPQKGHAVNLLDVPVPVARKLSAREQRDCEVIERLIKSYFLIVRKNIQDSVPKAVMHFLVNHVKDTLQSELVGQLYKSSLLDDLLTESEDMAQRRKEAADMLKALQGASQIIAEIRETHLW

種類予測

種類予測:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Pig

100

Horse

100

Bovine

100

Sheep

100

Dog

100

Rabbit

100

Zebrafish

Chicken

Xenopus

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - O00429 基板として

O00429

Site	PTM Type	Enzyme	Source
M1	Acetylation		Uniprot
S40	Phosphorylation	P49841 (GSK3B)	Uniprot
S44	Phosphorylation	P49841 (GSK3B)	Uniprot
T78	Phosphorylation		Uniprot
T79	Phosphorylation		Uniprot
K92	Ubiquitination		Uniprot
K97	Ubiquitination		Uniprot
K99	Ubiquitination		Uniprot
T102	Phosphorylation		Uniprot
R108	Methylation		Uniprot
K123	Ubiquitination		Uniprot
S126	Phosphorylation		Uniprot
K133	Ubiquitination		Uniprot
S136	Phosphorylation		Uniprot
K160	Ubiquitination		Uniprot
S179	Phosphorylation		Uniprot
T193	Phosphorylation		Uniprot
S200	Phosphorylation		Uniprot
K238	Ubiquitination		Uniprot
K255	Ubiquitination		Uniprot
K256	Ubiquitination		Uniprot
Y266	Phosphorylation		Uniprot
K271	Ubiquitination		Uniprot
K272	Ubiquitination		Uniprot
K283	Acetylation		Uniprot
S330	Phosphorylation		Uniprot
C361	S-Nitrosylation		Uniprot
C367	S-Nitrosylation		Uniprot
Y368	Phosphorylation		Uniprot
T394	Phosphorylation		Uniprot
T400	Phosphorylation		Uniprot
S412	Phosphorylation		Uniprot
Y449	Phosphorylation		Uniprot
S529	Phosphorylation		Uniprot
K532	Sumoylation		Uniprot
K535	Sumoylation		Uniprot
K535	Ubiquitination		Uniprot
S544	Phosphorylation		Uniprot
S548	Phosphorylation		Uniprot
S552	Phosphorylation		Uniprot
K558	Sumoylation		Uniprot
K568	Sumoylation		Uniprot
K568	Ubiquitination		Uniprot
S572	Phosphorylation		Uniprot
T586	Phosphorylation		Uniprot
K594	Sumoylation		Uniprot
T595	Phosphorylation		Uniprot
K597	Sumoylation		Uniprot
K597	Ubiquitination		Uniprot
K606	Sumoylation		Uniprot
S607	Phosphorylation		Uniprot
K608	Sumoylation		Uniprot
S616	Phosphorylation	P06493 (CDK1) , P28482 (MAPK1) , P27361 (MAPK3) , Q05655 (PRKCD) , Q00535 (CDK5) , P24941 (CDK2)	Uniprot
S637	Phosphorylation	P17612 (PRKACA) , Q13464 (ROCK1) , Q13131 (PRKAA1)	Uniprot
C644	S-Nitrosylation		Uniprot
S693	Phosphorylation	P49841 (GSK3B)	Uniprot
T701	Phosphorylation		Uniprot
S724	Phosphorylation		Uniprot

研究背景

O00429_DNM1L_HUMAN

機能:

Functions in mitochondrial and peroxisomal division. Mediates membrane fission through oligomerization into membrane-associated tubular structures that wrap around the scission site to constrict and sever the mitochondrial membrane through a GTP hydrolysis-dependent mechanism. The specific recruitment at scission sites is mediated by membrane receptors like MFF, MIEF1 and MIEF2 for mitochondrial membranes. While the recruitment by the membrane receptors is GTP-dependent, the following hydrolysis of GTP induces the dissociation from the receptors and allows DNM1L filaments to curl into closed rings that are probably sufficient to sever a double membrane. Through its function in mitochondrial division, ensures the survival of at least some types of postmitotic neurons, including Purkinje cells, by suppressing oxidative damage. Required for normal brain development, including that of cerebellum. Facilitates developmentally regulated apoptosis during neural tube formation. Required for a normal rate of cytochrome c release and caspase activation during apoptosis; this requirement may depend upon the cell type and the physiological apoptotic cues. Plays an important role in mitochondrial fission during mitosis. Required for formation of endocytic vesicles. Proposed to regulate synaptic vesicle membrane dynamics through association with BCL2L1 isoform Bcl-X(L) which stimulates its GTPase activity in synaptic vesicles; the function may require its recruitment by MFF to clathrin-containing vesicles. Required for programmed necrosis execution. Rhythmic control of its activity following phosphorylation at Ser-637 is essential for the circadian control of mitochondrial ATP production.

Inhibits peroxisomal division when overexpressed.

PTMs:

Phosphorylation/dephosphorylation events on two sites near the GED domain regulate mitochondrial fission. Phosphorylation on Ser-637 inhibits the GTPase activity, leading to a defect in mitochondrial fission promoting mitochondrial elongation. Dephosphorylated on this site by PPP3CA which promotes mitochondrial fission. Phosphorylation on Ser-616 activates the GTPase activity and promotes mitochondrial fission. Phosphorylated in a circadian manner at Ser-637.

Sumoylated on various lysine residues within the B domain, probably by MUL1. Sumoylation positively regulates mitochondrial fission. Desumoylated by SENP5 during G2/M transition of mitosis. Appears to be linked to its catalytic activity.

S-nitrosylation increases DNM1L dimerization, mitochondrial fission and causes neuronal damage.

Ubiquitination by MARCHF5 affects mitochondrial morphology.

O-GlcNAcylation augments the level of the GTP-bound active form of DRP1 and induces translocation from the cytoplasm to mitochondria in cardiomyocytes. It also decreases phosphorylation at Ser-637 (By similarity).

細胞の位置付け:

Cytoplasm>Cytosol. Golgi apparatus. Endomembrane system>Peripheral membrane protein. Mitochondrion outer membrane>Peripheral membrane protein. Peroxisome. Membrane>Clathrin-coated pit. Cytoplasmic vesicle>Secretory vesicle>Synaptic vesicle membrane.
Note: Mainly cytosolic. Translocated to the mitochondrial membrane through O-GlcNAcylation and interaction with FIS1. Recruited to the mitochondrial outer membrane by interaction with MIEF1. Colocalized with MARCHF5 at mitochondrial membrane. Localizes to mitochondria at sites of division. Localizes to mitochondria following necrosis induction. Associated with peroxisomal membranes, partly recruited there by PEX11B. May also be associated with endoplasmic reticulum tubules and cytoplasmic vesicles and found to be perinuclear. In some cell types, localizes to the Golgi complex. Binds to phospholipid membranes.

組織特異性:

サブユニット構造:

Homotetramer; dimerizes through the N-terminal GTP-middle region of one molecule binding to the GED domain of another DNM1L molecule. Oligomerizes in a GTP-dependent manner to form membrane-associated tubules with a spiral pattern. Interacts with GSK3B and MARCHF5. Interacts (via the GTPase and B domains) with UBE2I; the interaction promotes sumoylation of DNM1L, mainly in its B domain. Interacts with PPP3CA; the interaction dephosphorylates DNM1L and regulates its transition to mitochondria. Interacts with BCL2L1 isoform BCL-X(L) and CLTA; DNM1L and BCL2L1 isoform BCL-X(L) may form a complex in synaptic vesicles that also contains clathrin and MFF. Interacts with FIS1. Interacts with MIEF2 and MIEF1; GTP-dependent this regulates GTP hydrolysis and DNM1L oligomerization. Interacts with PGAM5; this interaction leads to dephosphorylation at Ser-656 and activation of GTPase activity and eventually to mitochondria fragmentation.

タンパク質ファミリー:

The GED domain folds back to interact, in cis, with the GTP-binding domain and middle domain, and interacts, in trans, with the GED domains of other DNM1L molecules, and is thus critical for activating GTPase activity and for DNM1L dimerization.

Belongs to the TRAFAC class dynamin-like GTPase superfamily. Dynamin/Fzo/YdjA family.

研究領域

· Cellular Processes > Cell growth and death > Necroptosis. (View pathway)

· Environmental Information Processing > Signal transduction > TNF signaling pathway. (View pathway)

· Organismal Systems > Immune system > NOD-like receptor signaling pathway. (View pathway)

参考文献

1). HSP70 attenuates compression-induced apoptosis of nucleus pulposus cells by suppressing mitochondrial fission via upregulating the expression of SIRT3. EXPERIMENTAL AND MOLECULAR MEDICINE, 2022 (PubMed: 35338257) [IF=12.8]

Application: IF/ICC Species: Rat Sample: NP cells

Fig. 2 HSP70 suppressed compression-induced apoptosis of NP cells. a, b The effects of TRC on apoptosis of NP cells measured by flow cytometry using Annexin V-FITC/PI staining. c Typical fluorescence photomicrograph of TUNEL staining of NP cells (scale bar: 50 μm). d, e The effects of VER on apoptosis of NP cells measured by flow cytometry using Annexin V-FITC/PI staining. f Typical fluorescence photomicrograph of TUNEL staining of NP cells (scale bar: 50 μm). (N = 3, **P < 0.01; ***P < 0.001).

Application: WB Species: Rat Sample: NP cells

Fig. 5 HSP70 suppressed compression-induced mitochondrial fission in NP cells. a The effects of TRC on the expression of DRP1, MFF, Fis1, Mfn1, Mfn2, and OPA1 (N = 3). b Representative fluorescence photomicrograph of TOM20 examined by immunofluorescence staining (N = 3, original magnification: ×1000). c The morphological ultrastructural appearance of mitochondria observed by TEM (N = 3). d The effects of VER on the expression of DRP1, MFF, Fis1, Mfn1, Mfn2, and OPA1 in NP cells treated with TRC (N = 3). e Representative fluorescence photomicrograph of TOM20 examined by immunofluorescence staining of NP cells treated with TRC and VER (N = 3, original magnification: ×1000). f Representative fluorescence photomicrograph of TOM20 examined by immunofluorescence staining of NP cells treated with TRC and HSP70-specific siRNAs (N = 3, original magnification: ×1000).

2). The nonstructural protein 1 of respiratory syncytial virus hijacks host mitophagy as a novel mitophagy receptor to evade the type I IFN response in HEp-2 cells. mBio, 2023 (PubMed: 37909764) [IF=6.4]

Application: WB Species: Human Sample:

Fig 2 RSV and RSV-NS1 protein could induce mitophagy by facilitating Drp1-dependent mitochondria fission. (A–C) HEp-2 cells were infected with RSV (MOI = 2) (A and C) or transfected with a HA-NS1 plasmid for 36 h (B). The mitochondrial morphology (A and B) and mitochondrial membrane potential detected by JC-1 staining (C) were observed by TEM and fluorescence microscope, respectively. Scale bar = 2.5 µm (A)/2.0 µm (B). (D–G) HEp-2 cells were infected with GFP-RSV (MOI = 2) (D and F) or transfected with HA-NS1 plasmids for 36 h (E and G), and then stained with MitoTracker Red CMXRos (D) or MitoTracker Green (E), respectively. Nuclei were stained with Hoechst. The mitochondrial morphology and relative expression of proteins were observed by laser confocal microscopy and WB. Scale bar = 20 µm. (H–J) HEp-2, HEp-2-NC, and HEp-2-DRP1-KD cells were infected with RSV (H and J) or transfected with a Flag-NS1 plasmid for 36 h (I). Relative expression of proteins and viral titers were detected by WB and viral plaque assay, respectively. Each data represents the mean ± SD of three independent experiments.

3). Valproic acid regulates MIEF1 through MST2-HIPPO to suppress breast cancer growth. Life sciences, 2022 (PubMed: 36126724) [IF=6.1]

4). Ginsenoside CK improves skeletal muscle insulin resistance by activating DRP1/PINK1-mediated mitophagy. Food & Function, 2023 (PubMed: 36562271) [IF=6.1]

5). FGFR2 Mutation p.Cys342Arg Enhances Mitochondrial Metabolism-Mediated Osteogenesis via FGF/FGFR-AMPK-Erk1/2 Axis in Crouzon Syndrome. Cells, 2022 (PubMed: 36231091) [IF=6.0]

Application: WB Species: Human Sample:

Figure 7 Mitochondrial dynamics participate in osteogenesis mediated by FGFR/FGFR2-AMPK pathway: (A) Mito-tracker red and mitochondria 2D analysis showed more fragmented or punctate mitochondria with fewer and shorter branches in the MT group, while there were widely reticular mitochondria with longer branches in the WT group. (B) qRT-PCR and Western blot analysis demonstrated that the expressions of mitochondrial-fusion-related factors Mfn2 and Opa1 were downregulated, however, the expression of mitochondrial-fission-related factor Drp1 was upregulated in the MT group compared to the WT group. (C) Mito-tracker red and mitochondria 2D analysis showed a filamentous network of mitochondria with more and longer branches widely spread in the cytoplasm in the siFGFR2 group. (D) qPCR and Western blot analysis showed that the expression of fusion-related genes Mfn2 and Opa1 was increased and the expression of fission-related gene Drp1 was decreased in the siFGFR2 group. (E) After treatment with Compound C, the morphology of the mitochondria which should have tended to split adopted a fused reticular structure with more and longer branches, as shown by Mito-tracker red and mitochondria 2D analysis. (F) Western blot analysis showed the level of MFN2 and OPA1 was increased and then level of DRP1 was decreased, as induced by Compound C. p values were significant at * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001.

6). Silibinin Induces G2/M Cell Cycle Arrest by Activating Drp1-Dependent Mitochondrial Fission in Cervical Cancer. Frontiers in Pharmacology, 2020 (PubMed: 32226384) [IF=5.6]

Application: WB Species: human Sample: cervical cancer cells

FIGURE 6 | SB impaired mitochondrial fission in cervical cancer cells. (B) The mitochondrial fission related protein expression extracted from cervical cancer cells. Drp1 expression was increased in both two cervical cancer cells. The mitochondrial fission related protein and gene expression level in cervical cancer cells were detected by western blotting and qPCR, respectively. Values (mean ± SDs) were obtained from at least three independent experiments.*P < 0.05, **P < 0.01, and ***P < 0.001 by one-way ANOVA with Tukey’s test.

7). Patchouli alcohol alleviates metabolic dysfunction-associated steatohepatitis via inhibiting mitochondria-associated endoplasmic reticulum membrane disruption-induced hepatic steatosis and inflammation in rats. International immunopharmacology, 2024 (PubMed: 38971107) [IF=5.6]

8). Baicalein inhibits mitochondrial apoptosis induced by oxidative stress in cardiomyocytes by stabilizing MARCH5 expression. JOURNAL OF CELLULAR AND MOLECULAR MEDICINE, 2020 (PubMed: 31880404) [IF=5.3]

Application: WB Species: mouse Sample: H9C2 cells

FIGURE 6|The cardioprotective pathway regulated by baicalein. H9C2 cells were exposed to 100μM H2O2 for 24 h after pre-treated with 50 μM baicalein for 4 h, transfected with MARCH5-cDNA or MARCH5-siRNA for 24 h, respectively. KLF4 and Drp1 proteins were detected by Western blotting. (A) The expression level of KLF4 and Drp1 in different time groups after treatment with H2O2

Application: WB Species: mouse Sample: H9C2 cells

FIGURE 6|The cardioprotective pathway regulated by baicalein. H9C2 cells were exposed to 100μM H2O2 for 24 h after pre-treated with 50 μM baicalein for 4 h, transfected with MARCH5-cDNA or MARCH5-siRNA for 24 h, respectively. KLF4 and Drp1 proteins were detected by Western blotting. (G) The ubiquitylation level of Drp1 in normal, MARCH5-siRNA and MARCH5-cDNA after treatment with H2O2.

9). Anemoside B4 alleviates arthritis pain via suppressing ferroptosis-mediated inflammation. Journal of cellular and molecular medicine, 2024 (PubMed: 38334255) [IF=5.3]

Application: WB Species: Mouse Sample:

FIGURE 5 (A) The molecular structure of AB4. (B–D) Molecular docking of AB4 with GSK‐3β. The modelled 3D structure of GSK‐3β docked with AB4 (B). The enlarged view of binding site in box (C). The interaction bonds of GSK‐3β with AB4 (D). Bonds showed as yellow dotted lines, and bond lengths were presented as numbers. (E) The titration between AB4 and GSK‐3β. The top panel presents typical calorimetric titration of AB4 with GSK‐3β at 25°C. The bottom panel shows the plots of the heat evolved (kcal) per mol of AB4 added corrected for the heat of with GSK‐3β, against the molar ratio of AB4 to GSK‐3β. Data solid squares were fitted to a single set of the identical sites model, and the solid line represented the best fit. (F) Representative immunofluorescence staining images of GSK‐3β and Drp1 in the spinal dorsal horn of the control, CIA and CIA + AB4 groups. Scale bar = 20 μm. (G) Quantitative analysis of the fluorescence intensity of GSK‐3β and Drp1. (H, I) Western blot analysis and quantitative grey value analysis of pGSK‐3β‐Tyr216, GSK‐3β, pDrp1‐Ser616, pDrp1‐Ser637 and Drp1 level in the spinal cord of the control, CIA and CIA + AB4 groups. Data are presented as mean ± SD (n = 5). *p

10). Evodiamine induces ROS-Dependent cytotoxicity in human gastric cancer cells via TRPV1/Ca2+ pathway. CHEMICO-BIOLOGICAL INTERACTIONS, 2022 (PubMed: 34808100) [IF=5.1]

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DRP1 Antibody - #DF7037

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